What are the disadvantages of tyramine amplification methods?

One main limitation of TSA is the potential for background signals. Non-specific binding of the tyramide-conjugated fluorophore or hapten can lead to background staining, reducing the signal-to-noise ratio and affecting the accuracy of target detection. TSA protocols also require optimization of various parameters such as tyramide concentration, reaction time, and amplification duration. Thus, achieving optimal conditions for each specific application can be time-consuming.